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Abstract: SA-OR50

Rhesus Macaque Serves as a Model for Human Lateral Branching Nephrogenesis

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 500 Development, Stem Cells, and Regenerative Medicine

Authors

  • Schuh, Meredith Posner, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
  • Alkhudairy, Lyan, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
  • Potter, Andrew, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
  • Potter, Steven, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
  • Chetal, Kashish, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
  • Salomonis, Nathan, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
  • Kopan, Raphael, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
Background

Premature infants are at risk for chronic kidney disease later in life due to low nephron endowment. Lateral branching nephrogenesis (LBN), not occurring in the mouse, is a poorly understood but critical period of human nephrogenesis. Here, we analyze third trimester LBN in the rhesus macaque at the molecular and morphological level.

Methods

The morphology of third trimester rhesus kidneys was assessed by immunostaining after tissue clearing. 3D renderings were created using Bitplane Imaris. Single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq) were performed on 4 kidneys from 4 rhesus using cold protease digestion and 10xGenomics platform. Unsupervised analyses using ICGS2 were used to identify distinct cell populations and GO-Elite was used to compare rhesus with human and mouse datasets.

Results

The gestational period of the rhesus lasts 165 days. We determined that cessation of rhesus nephrogenesis occurs between 136- and 138-days gestational age (GA). LBN was observed along the ureteric stalks at 126-138 days GA. We noted rosette-like patterns of ureteric tips and nephron progenitor cells (NPC) in both rhesus and human third trimester archival samples. scRNA-seq was performed on 4 cortically enriched rhesus samples 129-131 days GA revealing 37 transcriptionally distinct cell clusters. C25 was predicted to contain the naïve NPCs and included CITED1, MEOX1, and EYA1. snRNA-seq yielded 5,972 nuclei, corresponding to 29 ICGS2 clusters. We found a single cluster (c26), with a near identical GO-Elite enrichment profile to that the naïve NPC scRNA-seq cluster (c25). snRNA-seq c26 contained many unique markers not found in the matching scRNA-seq c25 (e.g., BMPER, DPP6, KIRREL3). GO-ELITE showed that late-gestation rhesus NPC markers more closely aligned to late-gestation murine NPCs, whereas the 2nd trimester human NPCs more closely aligned to mid-gestation murine NPCs. Novel surface markers predicted in the rhesus include CACNA1E, KIRREL3, and KCNB2.

Conclusion

The rhesus is the first animal model to demonstrate LBN, suggesting that LBN is conserved in old world primates. We identified novel genes upregulated during LBN and surface markers that could be used during cell-sorting. snRNA-seq of naïve NPC nascent transcripts may provide mechanistic insights that would otherwise be missed.

Funding

  • NIDDK Support